Functional Expression of a Single-chain Antibody Fragment Against Human Epidermal Growth Factor Receptor 2 (HER2) in Escherichia Coli

Overview

Journal J Ind Microbiol Biotechnol

Publisher Oxford University Press

Specialty Biotechnology

Date 2014 Mar 28

PMID 24671570

Citations 15

Authors

Vajihe Akbari

Hamid Mir Mohammad Sadeghi

Abbas Jafrian-Dehkordi

Daryoush Abedi

C Perry Chou

Affiliations

Soon will be listed here.

Abstract

The human epidermal growth factor receptor (HER) family plays an important role in cell growth and signaling and alteration of its function has been demonstrated in many different kinds of cancer. Receptor dimerization is necessary for the HER signal transduction pathway and tyrosine kinase activity. Recently, several monoclonal antibodies have been developed to directly interfere with ligand-HER receptor binding and receptor dimerization. A single chain variable fragment (ScFv) is a valuable alternative to an intact antibody. This report describes the production and purification of an ScFv specific for domain II of the HER2 receptor in Escherichia coli BL21 (DE3) cytoplasm. The majority of expressed of anti-her2his-ScFv protein was produced as inclusion bodies. A Ni-NTA affinity column was used to purify the anti-her2his-ScFv protein. The molecular weight of anti-her2his-ScFv protein was estimated to be approximately 27 kDa, as confirmed by SDS-PAGE and Western blotting assay. The anti-her2his-ScFv showed near 95 % purity and reached a yield of approximately 29 mg/l in flask fermentation. The purified anti-her2his-ScFv showed its biological activity by binding to HER2 receptor on the surface of BT-474 cells. This ScFv may be a potential pharmaceutical candidate for targeting tumour cells overexpressing HER2 receptor.

Citing Articles

Integrated bioprocessing and genetic strategies to enhance soluble expression of anti-HER2 immunotoxin in E. Coli.

Mani S, Arab B, Akbari V, Chou C AMB Express. 2024; 14(1):107.

PMID: 39341967 PMC: 11438746. DOI: 10.1186/s13568-024-01765-6.

Design and construction of scFv-PE35KDEL as a novel immunotoxin against human epidermal growth factor receptor 2 for cancer therapy.

Shariaty Vaziri Z, Shafiee F, Akbari V Biotechnol Lett. 2023; 45(4):537-550.

PMID: 36807722 DOI: 10.1007/s10529-023-03360-4.

Co-Expression of Chaperones for Improvement of Soluble Expression and Purification of An Anti-HER2 scFv in .

Estabragh A, Mohammad Sadeghi H, Akbari V Adv Biomed Res. 2023; 11:117.

PMID: 36798911 PMC: 9926028. DOI: 10.4103/abr.abr_351_21.

Therapeutic potential of a novel IP-10-(anti-HER2 scFv) fusion protein for the treatment of HER2-positive breast cancer.

Ahmadzadeh M, Mohit E Biotechnol Lett. 2023; 45(3):371-385.

PMID: 36650341 DOI: 10.1007/s10529-022-03342-y.

Different strategies for expression and purification of the CT26-poly-neoepitopes vaccine in Escherichia coli.

Movahed Z, Sharif E, Ahmadzadeh M, Nezafat N, Jahandar H, Mohit E Mol Biol Rep. 2022; 49(2):859-873.

PMID: 35059972 DOI: 10.1007/s11033-021-06727-w.

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